METHOD FOR EXTRACTING POLYSACCHARIDES FROM HIGHER PLANTS AND FUNGI THROUGH MICROWAVE CHEMICAL TREATMENT

• US 20140309414A1
• Filed: 11/01/2012
• Published: 10/16/2014
• Est. Priority Date: 11/07/2011
• Status: Active Grant

• Current Assignee: Shenyang Kesi High-Technology Co., Ltd.
• Original Assignee: Shenyang Kesi High-Technology Co., Ltd.
• Inventors: Zhang, Jinsong, Li, Mingtian, Liu, Zhiyu, Xu, Lei
• Granted Patent: US 9,447,198 B2
• Time in Patent Office: Days
• US Class Current: 536/123.1
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C08B 37/0003 General processes for their isolation or fractionation, e.g. purification or extraction from biomass
• CPC Class Codes: C08B 37/006 Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
• CPC Class Codes: C08H 8/00 Macromolecular compounds derived from lignocellulosic materials pretreatment thereof B27N

Abstract

This invention relates to a field of pharmaceutical chemistry, relates to a process of extracting high water soluble polysaccharides from higher plants or fungi. The present invention discloses a process of extracting higher plants and fungi polysaccharides based on a microwave chemistry method, comprising: putting the residue or pulverized higher plants and fungi after being degreased by an organic solvent into a microwave reaction chamber to react with an acid solution; and then distilling to remove excess acid or washing with organic solvent to remove the acid; adding water solution for extraction, subjecting the extracting solution after concentration to alcohol precipitation, separating precipitates aka polysaccharides therefrom.

The present invention has significant advantages like fast processing rate, high polysaccharides yield, low organic acid consumption and efficient and easy to recycle, low water consumption, low power consumption, etc., and obtained polysaccharides have high yield and purity, good water solubility, and good biological activity.

Claims

1. A process of extracting higher plants or fungi polysaccharides based on a microwave chemistry treatment, comprising the following steps:

a treating pulverized higher plants or fungi with an organic solvent to remove liposoluble components thereof, to obtain higher plants or fungi residue;

or directly using pulverized higher plants or fungi;

b) putting the residue or pulverized higher plants or fungi obtained in the step a) into a microwave reaction chamber, adding an acid solution of a mass concentration of 5% to 99%, conducting reaction of the mixer for 5-120 mins at a microwave power of mass power density of 1 kilowatt per kilogram of material-10 kilowatts per kilogram of material under a work pressure of 20 mmHg-760 mmHg;

optionally conducting concentration, and then washing with an organic solvent to remove excess acid; and

c) conducting extraction by adding water solution of 5-15 times by volume into the product obtained from the step b), subjecting the extraction solution after concentration to alcohol precipitation, separating precipitates, i.e. the polysaccharides.

2. The process of claim 1, wherein, the application method of the microwave power in said step b) is a continuous microwave mode or a combination of continuous microwave and pulse microwave modes;

wherein, in case of using the combination of continuous microwave and pulse microwave, the continuous microwave irradiation is first used until the acid solution refluxes, and then is switched to pulse microwave for 5 min-120 min.

3. The process of claim 1, wherein, in said step b), when the acid solution used is a non-volatile acid, removal of the acid by concentration is not conducted;

when volatile acid is used, concentration is first conducted to remove the acid, preferably by distillation under reduced pressure under microwave heating.

4. The process of claim 2, wherein, in said step b), in case of continuous microwave, mass power density is 1 kilowatt per kilogram of material-5 kilowatts per kilogram of material;

in case of pulse microwave, mass power density is 2 kilowatts per kilogram of material-10 kilowatts per kilogram of material, the duty ratio is A/B, where A=1 sec-100 sec, B=1 sec-100 sec.

5. The process of claim 2, wherein, the acid solution in said step b) is an organic acid or a mixed solution of an organic acid and an inorganic acid.

6. The process of claim 5, wherein, the organic acid solution in said step b) is selected from the group consisting of oxalic acid, formic acid, acetic acid, and propionic acid.

7. The process of claim 6, wherein, in said step b) a weight percentage concentration of oxalic acid is 5% to 50%, preferably 10%-35%;

a weight percentage concentration of formic acid is 10%-99%, preferably 30-85%;

a weight percentage concentration of acetic acid is 10%-99%, preferably 60-95%;

or a weight percentage concentration of propionic acid is 10%-99%, preferably 70-95%.

8. The process of claim 5, wherein, in the mixed solution of an organic acid and an inorganic acid used in said step b), the mass percentage concentration of the inorganic acid in the mixed solution is 0.1%-15%.

9. The process of claim 8, wherein, the inorganic acid in the step b) is selected from hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid.

10. The process of claim 1, wherein, in the step b), the ratio of the residue or pulverized higher plants or fungi obtained in step a) to the amount of the acid solution is 5/1-1/5.

11. The process of claim 1, wherein, the organic solvent used in said step b) is selected from methanol, ethanol, propanol or acetone.

12. The process of claim 1, wherein, the organic solvent used in said step a) is petroleum ether, methanol, ethanol, propanol or ethyl acetate.

13. The process of claim 1, wherein, the liposoluble component in the step a) is volatile oils, flavonoids, triterpenoids or saponins.

14. The process of claim 1, wherein, the alcohol used in said step c) is ethanol.

15. The process of claim 1, wherein, the higher plants is selected from flowers, leaves, seeds, barks, fruits, roots or tubers raw materials of astragalus, wolfberry, gingko leaf, papaya, honeysuckle, Chinese angelica, orange peel, ephedra sinica, ligusticum chuanxiong hort, acorns gramineus, garlic, sharpleaf galangal fruit, angelica, Chinese mugwort leaf, asarum, cistanche, elaeagnus angustifolia, eucalyptus leaf, cordata, ligustrum lucidum, notopterygium, ginseng, panax pseudoginseng, sarcandra glabra, plantago, polygonum orientale fruit, daphne genkwa, bergamot, white mulberry root-bark, mistletoe, scutellaria baicalensis, epimedium, tea leaf, rhodiola, aloe, oat, konjac, yam, gastrodia elata, radix bupleuri or acanthopanax;

the fungi is selected from fungal sporocarp or mycelium of lucid ganoderma, exidia auricula judae, mushrooms, polyporus, tremella, maitake, poria, rainbow conk, hericium erinaceus or cordyceps sinensis.

16. The process of claim 15, wherein, the higher plants is astragalus, wolfberry, yam, gingko leaf, panax pseudoginseng, plantago, gastrodia elata, eucommia ulmoides, salvia or kudzu;

the fungi is lucid ganoderma, poria, exidia auricula judae, or mushrooms.

17. The process of claim 1, wherein, the process of preparing astragalus polysaccharides comprising:

putting astragalus after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 10%-35% oxalic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of oxalic acid solution for 15-30 min, and then evaporating under reduced pressure the liquid in the microwave extraction chamber to dryness;

adding an ethanol solution of 3-5 times of the weight of the astragalus to the reaction chamber, stirring and washing for 40-60 minutes, and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration, combining the two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain astragalus polysaccharides.

18. An astragalus polysaccharide prepared using the process of claim 17, wherein, the molecular weight distribution of said astragalus polysaccharide is 3000-40000.

19. The process of claim 1, wherein, the process of preparing wolfberry polysaccharides, comprising:

putting wolfberry after being degreased by petroleum ether, ethanol into a microwave extraction chamber, adding 30%-85% formic acid solution of 1 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of formic acid solution for 15-25 min, and then evaporating under reduced pressure the formic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the wolfberry to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting the filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration, combining the two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain wolfberry polysaccharides.

20. A wolfberry polysaccharide prepared using the process of claim 19, wherein, the molecular weight distribution of said wolfberry polysaccharide is 3000-20000.

21. The process of claim 1, wherein, the process of preparing yam polysaccharides comprising:

putting yam after being pulverized into a microwave extraction chamber, adding 70%-95% propionic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of propionic acid solution for 15-25 min, and then evaporating under reduced pressure the propionic acid to dryness;

adding ethanol solution of 3-5 times of the weight of the yam to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration, combining the two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain yam polysaccharides.

22. A yam polysaccharides prepared using the process of claim 21, wherein the molecular weight distribution of said yam polysaccharide is 3000-20000.

23. The process of claim 1, wherein, the process of preparing gingko polysaccharides comprising:

putting gingko after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 30%-85% formic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of formic acid solution for 15-25 min, and then evaporating under reduced pressure the formic acid to dryness;

adding an ethanol solution of 4-6 times of the weight of the gingko to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration, combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain gingko polysaccharides.

24. A gingko polysaccharide prepared using the process of claim 23, wherein the molecular weight distribution of said gingko polysaccharide is 5000-12000.

25. The process of claim 1, wherein, the process of preparing panax pseudoginseng polysaccharides, comprising:

putting panax pseudoginseng after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 60%-95% acetic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of acetic acid solution for 15-25 min, and then evaporating under reduced pressure the acetic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the panax pseudoginseng to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration, combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain panax pseudoginseng polysaccharides.

26. A panax pseudoginseng polysaccharide prepared using the process of claim 25, wherein the molecular weight distribution of said panax pseudoginseng polysaccharide is 4000-20000.

27. The process of claim 1, wherein, the process of preparing plantago polysaccharides, comprising:

putting plantago after being degreased by petroleum ether, ethanol into a microwave extraction chamber, adding 80%-95% propionic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of propionic acid solution for 15-25 min, and then evaporating under reduced pressure the propionic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the plantago to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain plantago polysaccharides.

28. A plantago polysaccharide prepared using the process of claim 27, wherein the molecular weight distribution of said plantago polysaccharide is 4000-30000.

29. The process of claim 1, wherein, the process of preparing gastrodia elata polysaccharides comprising:

putting gastrodia elata after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding a formic acid-hydrochloride acid mixed solution of 0.3%-0.6% hydrochloride acid and 30%-85% formic acid of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of the acids mixed solution for 15-25 min, and then evaporating under reduced pressure the acids mixed solution to dryness;

adding an ethanol solution of 3-5 times of the weight of the gastrodia elata to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain gastrodia elata polysaccharides.

30. A gastrodia elata polysaccharide prepared using the process of claim 29, wherein the molecular weight distribution of said gastrodia elata polysaccharides is 3000-20000.

31. The process of claim 1, wherein, the process of preparing eucommia ulmoides polysaccharides comprising:

putting eucommia ulmoides after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 30%-85% formic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of formic acid solution for 15-25 min, and then evaporating under reduced pressure the formic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the eucommia ulmoides to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain eucommia ulmoides polysaccharides.

32. A eucommia ulmoides polysaccharide prepared using the process of claim 31, wherein the molecular weight distribution of said eucommia ulmoides polysaccharide is 3000-20000.

33. The process of claim 1, wherein, the process of preparing salvia polysaccharides comprising:

putting salvia after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 60%-95% acetic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of acetic acid solution for 15-25 min, and then evaporating under reduced pressure the acetic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the salvia to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain salvia polysaccharides.

34. A salvia polysaccharides prepared using the process of claim 33, wherein the molecular weight distribution of said salvia polysaccharides is 5000-25000.

35. The process of claim 1, wherein, the process of preparing kudzu polysaccharides comprising:

putting kudzu after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 70%-95% propionic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of propionic acid solution for 15-25 min, and then evaporating under reduced pressure the propionic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the kudzu to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain kudzu polysaccharides.

36. A kudzu polysaccharides prepared using the process of claim 35, wherein the molecular weight distribution of said kudzu polysaccharide is 3000-25000.

37. The process of claim 1, wherein, the process of preparing lucid ganoderma polysaccharides, comprising:

putting lucid ganoderma sporocarp after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 10%-35% oxalic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of oxalic acid solution for 15-25 min, and then evaporating under reduced pressure the liquid to dryness;

adding an ethanol solution of 3-5 times of the weight of the lucid ganoderma sporocarp to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain lucid ganoderma polysaccharides.

38. A lucid ganoderma polysaccharide prepared using the process of claim 37, wherein the molecular weight distribution of said lucid ganoderma polysaccharide is 3000-12000.

39. The process of claim 1, wherein, the process of preparing poria polysaccharides comprising:

putting poria after being pulverized and being degreased by ethanol into a microwave extraction chamber, adding 30%-85% formic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of formic acid solution for 15-25 min, and then evaporating under reduced pressure the formic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the poria to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain poria polysaccharides.

40. A poria polysaccharide prepared using the process of claim 39, wherein the molecular weight distribution of said poria polysaccharides is 2000-8000.

41. The process of claim 1, wherein, the process of preparing exidia auricula judae polysaccharides comprising:

putting exidia auricula judae after being pulverized into a microwave extraction chamber, adding 60%-95% acetic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of acetic acid solution for 15-25 min, and then evaporating under reduced pressure the acetic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the exidia auricula judae to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain exidia auricula judae polysaccharides.

42. An exidia auricula judae polysaccharides prepared using the process of claim 41, wherein the molecular weight distribution of said exidia auricula judae polysaccharide is 5000-40000.

43. The process of claim 1, wherein, the process of preparing mushroom polysaccharides comprising:

putting mushroom sporocarp after being pulverized into a microwave extraction chamber, adding 70%-95% propionic acid solution of 1.5 to 2.5 times of the weight thereof, at a microwave power density 1-2 KW/Kg under 500 mmHg-760 mmHg pressure maintaining the reflux of propionic acid solution for 15-25 min, and then evaporating under reduced pressure the propionic acid to dryness;

adding an ethanol solution of 3-5 times of the weight of the mushroom to the reaction chamber, stirring and washing for 40-60 minutes and conducting filtration;

extracting filtration residue after being dried twice with water, wherein each time the amount of water is 6-8 times of the weight of the residue, the extraction temperature is 70°

C., and the extraction time is about 40 minutes;

conducting filtration and combining two filtrate solutions, and concentrating the filtrate solution to a volume of ⅕

of the original extracting solution;

then adding ethanol for precipitation, and drying precipitates to obtain mushroom polysaccharides.

44. A mushroom polysaccharide prepared using the process of claim 43, wherein the molecular weight distribution of said mushroom polysaccharide is 3000-15000.